title: Ion selectivity and competition in channelrhodopsins.
abstract: Channelrhodopsins are light-gated ion channels of green algae. They are widely used for the analysis of neuronal networks using light in the emerging field of optogenetics. Under steady-state light conditions, the two open states, O1 and O2, mediate the photocurrents with different ion conductance and selectivity. To understand the conducting process as well as its optogenetic applications, it is important to study ion binding and transport of this promiscuous cation channel. Here, we present an enzyme kinetic algorithm that allowed us to calculate the ion composition of the initial and steady-state photocurrents for multication media. The approach is based on current-voltage relations determined for the individual ions H(+), Na(+), Ca(2+), and Mg(2+). We identify and quantify the widely different competition of the ions in wild-type channelrhodopsin-2 and two high-performing channelrhodopsin variants CatCh+ and C1V1. Both variants show enhanced Ca(2+) conductance, but only CatCh+ displays high steady-state Ca(2+) currents at neutral pH due to reduced H+ competition and low inactivation. We demonstrate that for optogenetic applications, one should always take into account that the variable equilibria of the two open states depend on light intensity, voltage, and the ionic composition of the medium.
authors.affiliation: Institute of Biology, Experimental Biophysics, Humboldt-Universität zu Berlin, Berlin, Germany. schneifr@hu-berlin.de
authors.forename: Franziska
authors.initials: F
authors.lastname: Schneider
authors.forename: Dietrich
authors.initials: D
authors.lastname: Gradmann
authors.forename: Peter
authors.initials: P
authors.lastname: Hegemann
mesh_terms: D001667:Binding, Competitive; D000075402:Channelrhodopsins; D004553:Electric Conductivity; D057809:HEK293 Cells; D006801:Humans; D015640:Ion Channel Gating / Q000528:radiation effects; D008027:Light; D008670:Metals / Q000378:metabolism*; D008954:Models, Biological; D011522:Protons; D013379:Substrate Specificity
chemical_list: D000075402:Channelrhodopsins; D008670:Metals; D011522:Protons